Methods of skin treatment and use of water-soluble beta-(1,3) glucans as active agents for producing therapeutic skin treatment agents

ABSTRACT

A method of treating an ulcer comprising applying to the ulcer a preparation comprising a water-soluble β-(1,3) glucan with β-(1,6) linked side-chains, where the side-chains comprise β-(1,3) linkages or up to four consecutive β-(1,6) linkages as active ingredients.

REFERENCE

The present application is a continuation-in-part of U.S. applicationSer. No. 09/936,748, filed on Sep. 12, 2001, which is incorporated byreference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention relates to the use of specific water soluble β-(1,3)glucans as active agents for combating the formation of wrinkles in theskin as well as epithelial tissue diseases.

2. Description of the Prior Art

The formation of wrinkles caused by increasing age is induced throughthe degradation of different macro molecules such as for example elastinand collagen, which are responsible for the elastases. Many inflammatoryskin diseases, such as for example psoriasis or UV erythema, can also becausatively be linked to an increased concentration of serine proteases,such as e.g. elastase in the upper skin areas (see R. Voegeli et al1996. in Cosm. Toil. 111, 51).

The formation of wrinkles in the skin is normally not counteracted bymeans of physiological active principles, but by means of cosmeticagents. Many so-called “anti-aging products” contain liposomes loadedwith water or aqueous active agents, which through the fat layer of theskin are reaching the epidermis, where they gradually dissolve andthrough continuous water release compensate the skin recesses andregulate the moisture content of the skin. However, this effect is nocombat against the causes, but only has a so-called “repairing effect”,which only lasts for a short period of time.

In contrast to this pure cosmetic use, cytostatic active agents are e.g.used for the abatement of psoriasis, such as selenium sulfide, cadmiumsulfide, zinc pyrithion or corticosteroid, the medical effect of whichresides e.g. in a reduction of the mitose activity in the basalmembrane. However, because of the known side effects these substancesshould not be used over extended periods of time. Further it is possibleto alleviate, but not heal, psoriasis by means of antiseptic activeagents, such as for example selenium oxide, salicylic acid, pyrithionederivatives, hexachlorophene or quaternary ammonium compounds or bymeans of cell dissolving and fat removing active substances such as forexample benzoy peroxide or tar extracts.

Ulcers manifest as lesions or sores in the skin or mucous membranesresulting from a successive disintegration of surface epithelial tissuesthat make them distinctive from wounds. The ulcer might be superficial,but in most cases the ulcer extends into deeper layers forming a cratersurrounded by defined edges, and is typically very painful. Theetiologies of the different ulcers are different, where faulty bloodcirculation, infection, nerve damage, trauma or cancers might beinductive. Examples of ulcers are diabetic ulcers, plantar ulcers,peptic ulcers, decubitus ulcers, venous ulcers, ischemic ulcers,cancerous ulcers, aphthous ulcers, sublingual ulcers, and atonic ulcers.Whereas, diabetic ulcers are known to have a very complicated etiology,many of the other ulcers have single or defined causes, whereininfections, faulty blood circulation and cancer would be directlyinvolved in the formation of apthous, decubitus, and cancerous ulcers,respectively. An ulcer is considered chronic when it does not heal in atimely fashion.

Diabetic ulcers represent a serious problem because of their delayed ordeficient healing often leading to chronic wounds that become high riskfor major complications including infections and amputations. The latteris substantiated by the fact that diabetic patients have the highestamputation rate of any type of chronic wound, in some prospectivestudies as much as 20% of the patients, adding to another 20%experiencing failed healing (see Margolis et al 1999, Diabetes Care22:692-695). Whereas, the etiology of most wounds and ulcers isidentified, that supports the development of preventive care and woundtreatment methods and drugs, this is not the case with diabetic ulcerswhere numerous factors contributing to the chronic wound development areinvolved. This includes predisposition for atherosclerosis andneuropathy, renal failure, microvascular disease, oedema, preponderancefor infections, impaired leukocyte function, increased destruction ofgrowth factors due to increased amounts of matrix metalloproteinases inthe wound fluid, and hyperglycemia. In addition, it has beendemonstrated that a macrophage dysfunction contributes to the disturbedwound healing process in diabetic ulcers (see Zykova et al. 2000,Diabetes 49:1451-1458). Several approaches to enhance the wound healingprocess in diabetic ulcers have been employed including the use oftransplants and growth factors, but so far with scarcely any success(see Greenhalgh 2003, Clin Plast Surg 30: 37-45).

The use of specific polysaccharides as agents against the skin aging isalso known from prior art. It has for example been proposed in U.S. Pat.No. 5,223,491 to use a carboxymethylated β-1,3 glucan, which has beenextracted from the yeast fungi Saccharomyces cerevisiae, for topicaluse. However, the glucan is insoluble in water and can therefore only beformulated with large difficulties.

In the European patent application EP-A1 0463540 (Taito) the use ofglucans against viruses is described. According to the teachings in thetwo papers DE-A1 3744345 (Lomapharm) and EP-B1 0175667 (Larm) glucansare only suited for stimulation of the activity of macrophages. Thepharmaceutical effect of different glucans is further known from the twoEuropean patent applications EP-A1 045338 (Debat) and EP-A1 0561408(Kaken). The object of the European patent EP-B1 0500718 (Donzis) is theuse of water insoluble β-(1,3) glucans, which have been obtained fromthe cell walls of yeasts, for revitalisation of the skin.

Also known from the prior art are very different solutions known forsmoothing of the skin and strengthening of the barrier function from acosmetic or medical view, which only solve a part of the problem andwhich may have strong side effects. Especially reference is made to theinternational patent application WO 98/40082 (Henkel), wherein the useof water soluble β-(1,3) glucans as active agents for the skin treatmentis described. These glucans, which preferably are schizopyhallan orkrestin, i.e. extracts of fungi, have in practice not shown to besufficiently effective. The complex task of the invention was thereforeto provide active agents which could be used against formation ofwrinkles in the skin (cosmetic effect) as well as skin diseases such as,for example, cradle cap, psoriasis, seborrheic dermatitis, seborrheasicca, seborrhea oleosa, psoriasis vulgaris, ichtyoses, UV erythemas, orulcers as exemplified by diabetes ulcer and aptheous ulcer(medicaleffect), and which can be both dermatological and toxicologicaltolerated and which improve the prior art, as described in WO 94/40082

SUMMARY OF THE INVENTION

A method for treating skin with a specific type of branched watersoluble beta-1,3-glucan is described. A defined water solublebeta-glucan is used to promote healing in ulcers as exemplified bydiabetic and aphthous ulcers. The soluble beta-glucan can be applied tothe affected epithelial layer area directly and when appropriate coveredby a gauze pad, or it can be used in various pharmaceutical formulationslike rinses, mixtures, gels, ointments, creams, or in combination withother appliances or agents.

DESCRIPTION OF THE FIGURE

FIG. 1: Individual curves for control (broken) and soluble beta-glucan(continuous) patients displayed as percentages of the ulcer areasrelative to areas at onset of study. The six patients in the controlgroup are denoted Ctr 1-6, while the nine patients receivingexperimental treatment with beta-glucan are denoted Glu 1-9.

DETAILED DESCRIPTION OF THE INVENTION

The object of the invention is the use of water soluble β-(1,3) glucans,which are substantially free from (1,6) linkages, as active agents forpreparation of therapeutical agents for skin treatment and skinvitalization, especially for manufacturing of agents which at the sametime work against skin aging and formation of wrinkles erythemas andwhich at the same time stimulate the growth of cells.

It was surprisingly found that water soluble β-glucans, whichpractically do not have (1,6) linkages, in the Langerhans cells in thedeeper skin layers initiate an immuno modulation, whereby the specialcytokines are produced and which are significantly superior to the knownglucans of the prior art (according to WO 98/40082), which have asignificant amount of (1,6) linkages.

Furthermore, it was surprisingly found that water soluble β-glucanapplied onto diabetic ulcers gave significant healing efficacy ascompared to best conventional wound care. Patients treated with solublebeta-glucan experienced complete healing of the diabetic ulcers during aperiod of 6 months, whereas only 30% of those receiving conventionalwound care experienced healing of the ulcers. Likewise it wassurprisingly found that the water soluble beta-glucan gave significanthealing efficacy to aphthous ulcers, a condition in which very limitedadequate treatments exist.

Water Soluble β-(1,3) Glucans

The term glucans is intended to mean homopolysaccharides based onglucose. Depending on steric linking there is a difference betweenβ-(1,3), β-(1,4) and β-(1,6) glucans. β-(1,3) Glucans normally show ahelical structure, whereas glucans with a (1,4) linkage generally have alinear structure. The β-glucans of the invention have a (1,3) structure,i.e. they are substantially free from undesired (1,6) linkages.Preferably, such β-(1,3) glucans are used where the side chainsexclusively show (1,3) linkages. Especially the agents contain glucanswhich are obtained on the basis of yeast from the family Sacchaomyces,preferably, Saccharomyces cerevisiae. Glucans of this type are utilizedin the present invention and are available in technical amountsaccording to known methods. The international patent application WO95/30022 (Biotec-Mackzymal) describes e.g. a method for producing suchsubstances, wherein glucans with β-(1,3) and β-(1,6) linkages arebrought in contact with β-(1,6) glucanases in such a way, thatpractically all β-(1,6) linkages are cleaved. More specifically, WO95/30022 states that the disclosed method ensures that most chains ofmore than four β-(1,6)-bound glucose units are cleaved off. Moreover,the enzyme treatment will only cleave β-(1,6)-linkages withinβ-(1,6)-linked chains, but will not remove the β-(1,6)-linked glucosylresidue extending from the branching points. The resulting enzymetreated glucan can be characterized as a branched β-(1,3)-glucan withβ-(1,3)-linked sidechains being attached by a β-(1,6)-linkage and beingessentially free of β-(1,6)-linked chains. The cleavage allows for up tofour consecutive β-(1,6) linkages or zero—four consecutive β(1,6)linkages. Preferably used for the manufacture of these glucans areglucanases based on Trichoderma harzianum.

As to the manufacture and availability of the glucans contained in theseagents, reference is made to the above-cited publication.

Commercial Applicability

For the purpose of the invention the water soluble β-(1,3) glucans finduse as active agents for manufacturing cosmetic and/or pharmaceuticalpreparations. Typical examples of such agents are skin care agents suchas for example anti-wrinkling cremes, anti-cellulitis cremes or sunprotection lotions as well as ointments for treating skin diseases suchas for example cradle cap, psoriasis, seborrheic dermatitis, seborrheasicca, seborrhea oleosa, psoriasis vulgaris, ichtyoses, UV erythemas ordiabetes ulcers. Normally the water soluble β-glucans can be used inamounts of 0.1 to 25, preferably 0.5 to 15 and especially 1 to 5% byweight, based on the agents. The agents can further as additionalauxiliary and additional agents contain mild surfactants, oil bodies,emulsifiers, hyperfatting agents, pearl lustre waxes, consistencysubstances, thickening agents, polymers, silicon compounds, fats, waxes,stabilizing agents, biogenic active substances, deodorants, agentsagainst dandruff, film forming agents, swelling agents, UV lightprotection factors, antioxidants, inorganic colour pigments,hydrotropes, preservatives, insect repellents, self tanning agents,solubilizing agents, perfume oils, colouring agents and the like.

The water soluble beta-glucan is used to promote the healing of chroniculcers, and various types of ulcers caused by poor blood circulation,infection, nerve damage, trauma or cancer, for example, diabetic ulcers,plantar ulcers, peptic ulcers, decubitus ulcers, venous ulcers, ischemiculcers, cancerous ulcers, aphthous ulcers, sublingual ulcers and atomiculcers. When the water soluble beta-glucan is utilized in treatingulcers, it may be present in a composition in amounts ranging from about0.1 to about 25%, based on the weight of the ingredients in thecomposition as recited above.

The soluble beta-glucan can be applied directly to the affected area asa mixture, or rinse, or gel followed by e.g. a dressing or a gauze padwhen appropriate, or used in combination with pharmaceutical applianceslike bandages, dressings, artificial skins, liposome or micelleformulations, microcapsules, aqueous vehicles for soaking gauzedressings, and the like, and mixtures thereof. Furthermore, the solublebeta-glucan can be applied by such as creams, gels, formulations, foams,ointments and sprays, salves, and films, which are intended to beapplied to the affected area.

Typical examples of suitable mild, i.e. especially skin compatiblesurfactants, are fatty alcohol polyglycol ether sulphates, monoglyceridesulphates, mono- and/or dialkyl sulfosuccinates, fatty acidisethionates, fatty acid sarcosinates, fatty acid taurides, fatty acidglutamates, α-olefine sulphonates, ethercarboxylic acids, alkyloligoglucosides, fatty acid glucamides, alkylamido betaines and/orprotein fatty acid condensates, the last mentioned preferably based onwheat proteins.

As oil bodies use can be made of, for example, Guerbet alcohols based onfatty alcohols with 6 to 18, preferably 8 to 10 carbon atoms, esters oflinear C₆-C₂₂ fatty acids with linear C₆-C₂₂ fatty alcohols, esters ofbranched C₆-C₁₃ carboxylic acids with linear C₆-C₂₂ fatty alcohols, suchas e.g. myristyl myristate, myristyl palmitate, myristyl stearate,myristyl isostearate, myristyl oleate, myristyl behenate, myristylerucate, cetyl myristate, cetyl palmitate, cetyl stearate, cetylisostearate, cetyl oleate, cetyl behenate, cetyl erucate, stearylmyristate, stearyl palmitate, stearyl stearate, stearyl isostearate,stearyl oleate, stearyl behenate, stearyl erucate, isostearyl myristate,isostearyl palmitate, isostearyl stearate, isostearyl isostearate,isostearyl oleate, isosteayl behenate, isostearyl oleate, oleylmyristate, oleyl palmitate, oleyl stearate, oleyl isostearate, oleyloleate, oleyl behenate, oleyl erucate, behenyl myristate, behenylpalmitate, behenyl stearate, behenyl isostearate, behenyl oleate,behenyl behenate, behenyl erucate, erucyl myristate, erucyl palmitate,erucyl stearate, erucyl isostearate, erucyl oleate, erucyl behenate anderucyl erucate. In additon esters of linear C₆-C₂₂ fatty acids withbranched alcohols, especially 2-ethylhexanol, esters ofhydroxycarboxylic acids with linear or branched C₆-C₂₂ fatty alcohols,especially dioctyl malate, esters of linear and/or branched fatty acidswith polyvalent alcohols (such as e.g. propylene glycol, dimeric diol ortrimeric triol) and/or Guerbet alcohols, triglycerides based on C₆-C₁₀fatty acids, liquid mixtures of mono-/di-/triglycerides based on C₆-C₁₈fatty acids, esters of C₆-C₂₂ fatty alcohols and/or Guerbet alcoholswith aromatic carboxylic acids, especially benzoic acid, esters ofC₂-C₁₂ dicarboxylic acids with linear or branched alcohols with 1 to 22carbon atoms or polyols with 2 to 10 carbon atoms and 2 to 6 hydroxylgroups, plant oils, branched primary alcohols, substituted cyclohexanes,linear and branched C₆-C₂₂ fatty alcohol carbonates, Guerbet carbonates,esters of benzoic acid with linear and/or branched C₆-C₂₂ alcohols (e.g.Finsolv® TN), linear or branched, symmetrical or unsymmetrical dialkylethers with 6 to 22 carbon atoms in each alkyl group, ring openingproducts of epoxydated fatty acid esters with polyols, silicone oilsand/or aliphatic or naphthenic hydrocarbons, such as e.g. squalan,squalen or dialkyl cyclohexanes, can be used

As emulsifiers, for example, nonionic surfactants from at least one ofthe following groups may be used:

-   -   (1) Addition products of 2 to 30 moles ethylene oxide and/or 0        to 5 moles propylene oxide on linear fatty alcohols with 8 to 22        C atoms, on fatty acids with 12 to 22 C atoms and on alkyl        phenols with 8 to 15 C atoms in the alkyl group;    -   (2) C_(12/18) fatty acid mono- and -diesters of addition        products of 1 to 30 moles ethylene oxide and glycerol;    -   (3) glycerol mono- and diesters and sorbitan mono- and diesters        of saturated and unsaturated fatty acids with 6 to 22 carbon        atoms and their ethylene oxide addition products;    -   (4) alkyl mono- and oligoglycosides with 8 to 22 carbon atoms in        the alkyl group and their ethoxylated analogues;    -   (5) addition products of 15 to 60 moles ethylene oxide on        ricinus oil and/or hardened ricinus oil;    -   (6) polyol and especially polyglycerol esters, such as e.g.        polyglycerol polyricinoleate, polyglycerol        poly-12-hydroxystearate or polyglycerol dimerate isostearate,        and also mixtures of compounds from more of these classes of        substances;    -   (7) addition products of 2 to 15 moles ethylene oxide on ricinus        oil and/or hardened ricinus oil;    -   (8) partial esters based on linear, branched, unsaturated or        saturated C_(6/22) fatty acids, ricinolic acid and 12-hydroxy        stearic acid and glycerol, polyglycerol, pentaerythrite,        dipentaerythrite, sugar alcohols (e.g. sorbitol), alkyl        glucosides (e.g. methyl glucoside, butyl glucoside, lauryl        glucoside) as well as polyglucosides (e.g. cellulose);    -   (9) mono-, di- and trialkylphosphates as well as mono-, di-        and/or tri-PEG alkylphosphates and their salts;    -   (10) wool wax alcohols;    -   (11) polysiloxane/polyalkyl/polyether copolymers or        corresponding derivatives;    -   (12) mixed esters of pentaerythrite, fatty acids, citric acid        and fatty alcohol according to DE 1165574 PS and/or mixed esters        of fatty acids with 6 to 22 carbon atoms, methyl glucose and        polyols, preferably glycerol or polyglycerol,    -   (13) polyalkylene glycols, as well as    -   (14) glycerol carbonate.

The addition products of ethylene oxide and/or of propylene oxide onfatty alcohols, fatty acids, alkyl phenols, glycerol mono- and diestersas well as sorbitan mono- and -diesters of fatty acids or on ricinus oilare known products which are commercially available. They are mixturesof homologous substances, with average degree of alkoxylationcorresponding to the ratio of the amounts of the substances ethyleneoxide and/or propylen oxide and substrate, with which the additionreaction is carried out. C_(12/18) fatty acid mono- and -diesters ofaddition products of ethylene oxide on glycerol are known from DE2024051 PS as revertive fatting agents for cosmetic preparations.

C_(8/18) alkyl mono- and oligoglycosides, their manufacture and theiruse is known from prior art. Their preparation can especially be carriedout by reaction of glucose or oligosaccharides with primary alcoholshaving 8 to 18 C atoms. With regard to the glycoside residue bothmonoglycosides, where a cyclic sugar group is glycosidic bond to thefatty alcohol, and oligomeric glycosides with a degree ofoligomerisation until preferably about 8, are suitable. The degree ofoligomerization is then a statistical mean value, based on adistribution of homologues which is usual for such products of technicalquality.

Zwitterionic surfactants can also be used as emulsifiers. The termzwitterionic surfactants is intended to mean such surface activecompounds which in their molecule have at least a quatenary ammoniumgroup and at least one carboxylate and one sulphonate group. Especiallysuitable zwitterionic surfactants are the so-called betaines such as theN-alkyl-N,N-dimethyl ammonium glycinates, for example the cocoalkyldimethyl ammonium glycinate, N-acylaminopropyl-N,N-dimethylammonium glycinate, for example the coco acylaminopropyl dimethylammonium glycinate, and 2-alkyl-3-carboxylmethyl-hydroxyethylimidazoline with in each case 8 to 18 C atoms in the alkyl oracyl-groups, as well as the coco acylaminoethylhydroxyethylcarboxymethyl glycinate. Especially preferred is that underthe CTFA term cocamidopropyl betaine known fatty acid amide derivative.Also suitable emulsifiers are ampholytic surfactants. Ampholyticsurfactants are such surface active compounds which in addition to aC_(8/18) alkyl or acyl group in the molecule at least contain a freeamino group and at least one —COOH or —SO₃H group and which can forminner salts. Examples of suitable ampholytic surfactants are N-alkylglycines, N-alkyl propionic acids, N-alkyl aminobutyric acids, N-alkyliminodipropionic acids, N-hydroxyethyl-N-alkylamidopropyl glycines,N-alkyltaurines, N-alkylsarcosines, 2-alkylaminopropionic acids andalkylamino acetic acids with in each case about 8 to 18 C atoms in thealkyl group. Especially preferable ampholytic surfactants are the N-cocoalkylamino propionate, the coco acylamino ethylaminopropionate and theC_(12/18) acylsarcosine. In addition to the ampholytic, also quaternaryemulsifiers can be used, of which ester salts of the type of esterquats,preferably methylquaternised di-fatty acid triethanolamine ester salts,are especially preferable.

As hyperfatting agents substances such as for example lanolin andlecithin as well as polyethoxylated or acylated lanolin and lecithinderivatives, polyol fatty acid esters, monoglycerides and fatty acidalkanolamides can be used, whereby the last mentioned at the same timeact as foam stabilisers.

As exemplary pearl gloss waxes the following should be mentioned:Alkylene glycolester, especially ethyleneglycol distearate; fatty acidalkanolamides, especially coco fatty acid diethanolamide; partialglycerides, especially stearic acid monoglyceride; esters of polyvalent,possibly hydroxysubstituted carboxylic acids with fatty alcohols with 6to 22 carbon atoms, especially long chain esters of tartaric acid; fatsubstances, such as for example fatty alcohols, fatty ketones, fattyaldehydes, fatty ethers and fatty carbonates, wherin the sum of carbonatoms is at least 24, especially lauron and distearyl ethers; fattyacids such as stearic acid, hydroxystearic acid or behenic acid, ringopening products of olefine epoxides with 12 to 22 carbon atoms withfatty alcohols with 12 to 22 carbon atoms and/or polyols with 2 to 15carbon atoms and 2 to 10 hydroxyl groups as well as their mixtures.

As consistency givers preferably use is made of fatty alcohols orhydroxy fatty alcohols with 12 to 22 and preferably 16 to 18 carbonatoms and additionally partial giycerides, fatty acids or hydroxy fattyacids. A combination of these substances with alkyl oligoglucosidesand/or fatty acid-N-methyl glucamides with the same chain length and/orpolyglycerol-poly-12-hydroxy stearates.

Suitable thickening agents are for example types of aerosil (hydrophilicsilicic acids), polysaccharides, especially xanthan gum, guar-guar,agar-agar, alginates and tyloses, carboxymethyl celluloses andhydroxyethyl celluloses, as well as higher molecular polyethylenglycolmono- and diesters of fatty acids, polyacrylates, (e.g. Carbopols® fromGoodrich or Synthalenes® from Sigma), polyacrylamides, polyvinyl alcoholand polyvinyl pyrrolidone, surfactants such as for example ethoxylatedfatty acid glycerides, ester of fatty acids with polyols such as forexample pentaerythrite or trimethylolpropane, fatty alcohol ethoxytateswith narrow distribution of homologous or alkyl oligoglucosides as wellas electrolytes such as sodium chloride and ammonium chloride.

Suitable cationic polymers are for example cationic cellulosederivatives, such as e.g. a quaternized hydroxyethyl cellulose, which isavailable under the name of Polymer JR 400® from Amerchol, cationicstarch, copolymers of diallyl ammonium salts and acrylamides,quaternized vinylpyrrolidone/vinylimidazol polymers, such as e.g.Luviquat® (BASF), condensation products of polyglycols and amines,quaternized collagen polypeptides, such as for example lauryl dimoniumhydroxypropyl hydrolyzed collagen (Lamequat®L/Grünau), quaternized wheatpolypeptides, polyethyleneimine, cationic silicone polymers, such ase.g. amidomethicones, copolymers of adipic acid and dimethylaminohydroxypropyl diethylenetriamine (Cartaretine®/Sandoz), copolymers ofacrylic acid with dimethyl diallylammonium chloride (Merquat® 550/Chemviron), polyamino polyamides, such as e.g. described in FR 2252840A, as well as their cross-linked water soluble polymers, cationic chitinderivatives such as for example quaternized chitosane, possibly microcrystalline distributed, condensation products of dihalogen alkyls, suchas e.g. dibromobutane with bisdialkylamines, such as e.g.bis-dimethylamino-1,3-propane, cationic guar-gum, such as e.g. Jaguar®CBS, Jaguar® C-17, Jaguar® C-16 from Celanese, quaternised ammonium saltpolymers, such as e.g. Mirapol® A-15, Mirapol® AD-1, Mirapol® AZ-1 fromMiranol.

As exemplary anionic, zwitterionic, amphoteric and non-ionic polymersthe following can be used: Vinyl acetate/crotonic acid copolymers, vinylpyrrolidone/vinyl acrylate copolymers, vinyl acetate/butylmaleate/isobornyl acrylate copolymers, methyl vinylether/maleic acidanhydride copolymers and their esters, non-cross-linked and with polyolscross-linked polyacrylic acids, acrylamido propyltrimethyl ammoniumchloride/acrylate copolymers, octylacrylamide/methyl methacrylate/tert.-butylaminoethyl methacrylate/2-hydroxypropyl methacrylatecopolymers, polyvinylpyrrolidone, vinylpyrrolidone/ vinylacetatecopolymers, vinylpyrrolidon/ dimethylamino ethylmethacrylate/vinylcaprolactam terpolymers as well as possibly derivatized cellulose ethersand silicones.

Suitable silicon compounds are for example dimethyl polysiloxane,methylphenyl polysiloxane, cyclic silicones as well as amino, fattyacid, alcohol, polyether, epoxy, fluorine, glycoside and/or alkylmodified silicon compounds, which at room temperature can be in theliquid as well as in the resin state. Further suitable are simethicones,which are mixtures of dimethicones with an average chain length of 200to 300 dimethyl siloxane units and hydrogenated silicates. A detailedsurvey of suitable volatile silicones can also be found in Todd et al.,Cosm. Toil. 91, 27 (1976).

Typical exemplary fats are glycerides, and as waxes natural waxes amongothers, can be used, such as e.g. candelilla wax, carnauba wax, Japanwax, espartogras wax, cork wax, guaruma wax, rice seed oil wax, sugarcane wax, ouricury wax, montan wax, beeswax, schellack wax, spermaceti,lanolin (wool wax), burzel fat, ceresin, ozokerit (terrestrial wax),petrolatum, paraffin waxes, micro waxes; chemically modified waxes (hardwaxes), such as e.g. montanester waxes, sasot waxes, hydrogenated yoyobawaxes as well as synthetic waxes, such as e.g. polyalkylene waxes andpolyethylene glycol waxes.

As stabilizers metal salts of fatty acids, such as e.g. magnesium,aluminium and/or zinc stearate or ricinoleate can be used.

As biogenic active substances should be understood for exampletocopherol, tocopherol acetate, tocopherol palmitate, ascorbic acid,desoxy ribonucleic acid, retinol, bisabolol, allantoin, phytantriol,panthenol, AHA acids, aminoacids, ceramides, pseudoceramides, essentialoils, extracts of plants and vitamin complexes.

As deo active agents e.g. antiperspirants such as aluminiumchlorohydrate come into question. This agent is in the form ofcolourless, hygroscopic crystals, which easily melt in air, and isobtained through evaporation of solutions of aluminium chloride inwater. Aluminium chlorohydrate is used for manufacturing of perspirationinhibiting and deodorising preparations and has probably its effectthrough the partial closure of the perspiratory gland by means ofprecipitation of proteins and/or polysaccharides (see J. Soc. Cosm.Chem.24, 281 (1973)). Under the trade name Locron® of Hoechst AG,Frankfurt/FRG, an aluminium chlorohydrate is for example on the market,which corresponds to the formula [Al₂(OH)₅Cl].2.5 H₂O, and use of thisis especially preferred (see J.Pharm.Pharmacol. 26, 531 (1975)]. Inaddition to the chlorohydrates also aluminium hydroxylactates as well asacid aluminium/zirconium salts can be used. As further deo active agentsesterase inhibitors can be added. These are preferably trialkyl citratessuch as trihethyl citrate, tripropyl citrate, triisopropyl citrate,tributyl citrate and especially triethyl citrate (Hydagen® CAT, HenkelKGaA, Düsseldorf/FRG). The substances inhibit the enzyme activity andthereby reduce the formation of odours. Probably the free acid isthereby set free through the cleavage of the citric acid ester, and thisacid lowers the pH value of the skin so much that the enzymes therebyare inhibited. Further substances which can be used as estersaseinhibitors are sterol sulphates or phosphates, such as for examplelanosterol, cholesterol, campesterol, stigmasterol and sitosterolsulphate or phosphate, Dicarboxylic acids and their esters, such as forexample glutaric acid, glutaric acid monoethylester, glutaric aciddiethylester, adipic acid, adipic acid monoethylester, adipic aciddiethylester, malonic acid and malonic acid diethylester,hydroxycarboxylic acids and their esters, such as for example citricacid, malic acid, tartaric acid or tartaric acid diethylester.Antibacterial active substances, which influence the germ flora and killsweat destroying bacteria or inhibit their growth, can also be containedin the pin preparations. Examples of this are chitosan, phenoxyethanoland chlorohexidin gluconate. Also5-chloro-2-(2,4-dichlorophen-oxy)-phenol has shown to have an especiallygood effect, and this product is marketed unter the trade name Irgasan®by Ciba-Geigy, Basel/CH.

As anti-dandruff agents, climbazol, octopirox and zinc pyrethion can beused. Useable film formation agents are for example chitosan,microcrystalline chitosan, quaternary chitosan, polyvinylpyrrolidon,vinylpyrrolidon/vinylacetate copolymers, polymers of the acrylic acids,quaternary derivatives of cellulose, collagen, hyaluronic acid or itssalts and similar compounds. As swelling agents for aqueous phases,montmorillonite, clay mineral substances, pemulen, as well asalkylmodified Carbopol types (Goodrich) can be used. Further suitablepolymers or swelling agents can be found in the survey of R.Lochhead inCosm. Toil. 108, 95 (1993).

UV light protection factors are e.g organic substances (light protectionfilters) which by room temperature are in liquid or crystalline form,and which are capable of absorbing ultraviolet radiation and to set freethe received energy in the form of radiation with long wavelength, e.g.in the form of heat. UVB filters can be soluble in oils or in water. Asoil soluble substances the following are mentioned as examples:

-   -   3-benzyliden camphor, respectively 3-benzylidene norcamphor and        the derivatives thereof, e.g. 3-(4-methylbenzylidene) camphor as        described in EP-B1 0693471;    -   4-aminobenzoic acid derivatives, preferably 4-(dimethylamino)        benzoic acid 2-ethylhexylester, 4-(dimethylamino) benzoic acid        2-octylester and 4-(dimethylamino) benzoic acid amylester;    -   esters of cinnamonic acid, preferably 4-methoxy cinnamonic acid        2-ethylhexylester, 4-methoxy cinnamonic acid propylester,        4-methoxy cinnamonic acid isoamylester, 2-cyano-3,3-phenyl        cinnamonic acid 2-ethythexylester (octocrylene);    -   esters of salicylic acid, preferably salicylic acid        2-ethylhexylester, salicylic acid 4-isopropyl benzylester,        salicylic acid homomenthylester;    -   derivatives of benzophenone, preferably 2-hydroxy-4-methoxy        benzophenone, 2-hydroxy-4-methoxy-4′-methyl benzophenone,        2,2′-dihydroxy-4-methoxy benzophenone;    -   esters of benzalmalonic acid, preferably 4-methoxy benzmalonic        acid 2-ethylhexyl ester,    -   triazine derivatives, such as e.g.        2,4,6-trianilino-(p-carbo-2′-ethyl-1′-hexyloxy)-1,3,5-triazine        and octyltriazone, as described in EP A1 0818450;    -   propane-1,3-diones, such as e.g.        1-(4-tert.-butylphenyl)-3-(4′-methoxy-phenyl)-propane-1,3-dion;    -   ketotricyclo(5,2,1,0)-decane derivatives, as described in EP-B1        06945521.

As water soluble substances the following can be mentioned:

-   -   2-Phenylbenzimidazol-5-sulphonic acid and the alkali, alkaline        earth, ammonium, alkylammonium, alkanolammonium and glucammonium        salts;    -   sulphonic acid derivatives of benzophenones, preferably        2-hydroxy-4-methoxybenzophenon-5-sulphonic acid and their salts;    -   sulphonic acid derivatives of 3-benzylidencamphen, such as e.g.        4-(2-oxo-3-bornylidenmethyl)-benzene sulphonic acid and        2-methyl-5-(2-oxo-bornyliden) sulphonic acid and their salts.

As typical UV-A filters especially derivatives of benzoyl methane comesin question, such as e.g.1-(4′-tert.-butylphenyl)-3-(4′-methoxyphenyl)propane-1,3-dion,4-tert.butyl-4′-methoxydibenzoyl-methane (Parsol 1789), or1-phenyl-3-(4′-isopropylphenyl-propane-1,3-dion. The UV-A and UV-Bfilters can of course also be used in mixtures. In this casecombinations of octocrylene or camphor derivatives with butylmethoxydibenzoylmethane are especially photosensitive.

In addition to the mentioned soluble substances also insoluble lightprotection pigments can be used for this purpose, i.e. fine dispersemetal oxides or salts. Examples of suitable metal oxides are especiallyzinc oxide and titanium dioxide and in addition other oxides of iron,zirconium, silicon, manganese, aluminium and cerium, as well as theirmixtures. As salts silicates (TALL), barium sulphate or zinc stearatecan be used. The oxides and salts are used in the form of the pigmentsfor skin caring and skin protecting emulsions and decorative cosmetics.The particles should have an average diameter of less than 100 nm,preferably between 5 and 50 nm and especially between 15 and 30 nm. Theycan have a spherical shape, but particles can also be used which have anellipsoidal form or else have a shape which differs from the sphericalshape. In sun protecting agents preferably so-called micro or nanopigments are used. Preferably micronized zinc oxide is used. Furthersuitable UV light protection factors can be found in the survey byP.Finkel in SÖFW-Journal 12Z 543 (1996). Likewise suitable are herbalextracts with UV absorbing or antioxidative properties.

In addition to the primary light protection substances also secondarylight protection substances of the antioxidant type find use, whichinterrupt the photochemichal reaction chain, which is initiated when UVradiation penetrates the skin. Typical examples of such are amino acids(e.g. glycin, histidin, tyrosin, tryptophan) and their derivatives,imidazoles (e.g. urocaninic acid) and their derivatives, peptides suchas D,L-camosine, D-camosine, L-camosine and their derivatives (e.g.anserine), carotinoides, carotine (e.g. α-carotin, β-carotin, lycopin)and their derivatives, chlorogenic acid and its derivatives, liponicacid and its derivatives (e.g. dihydroliponic acid), aurothioglucose,propylthiouracil and other thiols (e.g. thioredoxin, glutathion,cystein, cystin, cystamine and their glycosyl, n-acetyl, methyl, ethyl,propyl, amyl, butyl and lauryl, palmitoyl, oleyl, γ-linoleyl,cholesteryl and glyceryl esters) as well as their salts, dilaurylthiodipropionate, distearyl thiodipropionate, thiodipropionic acid andtheir derivatives (esters, ethers, peptides, lipides, nucleotides,nucleosides and salts) as well as sulfoximine compounds (e.g. buthioninsulfoximines, homocystein sulfoximines, butionin sulfones, penta-,hexa-, hepta-thionin sufoximine) in very small compatible doses (e.g.pmol to μmol/kg), further (metal) chelating agents (e.g. α-hydroxy fattyacids, palmitic acid, phytinic acid, lactoferrine), α-hydroxy acids(e.g. citric acid, lactic acid, malic acid), humin acid, gallic acid,gallic extracts, bilirubin, bifiverdin, EDTA, EGTA and theirderivatives, unsaturated fatty acids and their derivatives (e.g.γ-linolenic acid, linolic acid, oleic acid), folic acid and theirderivatives, ubichinon and ubichinol and their derivatives, vitamin Cand derivatives (e.g. ascorbyl palmitate, Mg-ascorbyl phosphate,ascorbyl acetate), tocopheroles and derivatives (e.g. vitamin Eacetate), vitamin A and derivatives (vitamin A patmitate) as well askoniferyl benzoate of benzoe resin, rutinic acid and their derivatives,α-glycosylrutin, ferula acid, furfuryliden glucitol, carnosine,butylhydroxy toluene, butylhydroxy anisol, nordihydro guajak resin acid,nordihydro guajaret acid, trihydroxy butyrophenon, uric acid and theirderivatives, mannose and its derivatives, super oxide dismutase, zincand its derivatives (e.a. ZnO, ZnSO₄), selen and its derivatives (e.g.selen-methionin), stilbenes and their derivatives (e.g. stilben oxide,trans-stilben oxide) and the derivatives suitable according to theinvention (salts, esters, ethers, sugars, nucleotides, nucleosides,peptides and lipids) of these mentioned active substances.

For improvement of the flow properties further hydrotropes, such as forexample ethanol, isopropyl alcohol, or polyols can be used. Polyolswhich in this case can be used preferably have 2 to 15 carbon atoms andat least two hydroxyl groups. The polyols can further contain additionalfunctional groups, especially amino groups, or be modified withnitrogen. Typical examples are:

-   -   Glycerol;    -   alkylen glycols, such as for example ethylene glycol, diethylene        glycol, propylene glycol, butylene glycol, hexylene glycol as        well as polyethylen glycols with an average molecular weight        from 100 to 1 000 Daltons;    -   oligoglycerol mixtures of technical quality with a        self-condensation degree of 1.5 to 10, such as e.g. technical        quality diglycerol mixtures with a diglycerol content of 40 to        50% by weight;    -   methyol compounds, such as especially trimethylol ethane,        trimethylol propane, trimethylol butane, pentaerythrite and        dipentaerythrite;    -   low alkyl glucosides, especially such with 1 to 8 carbons in the        alkyl residue, such as for example methyl and butyl glucoside;    -   sugar alcohols with 5 to 12 carbon atoms, such as for example        sorbitol or mannit;    -   sugars with 5 to 12 carbon atoms, such as for example glucose or        saccharose;    -   aminosugars, such as for example glucamine;    -   dialcoholamines, such as diethanolamine or        2-amino-1,3-propanediol.

As preservatives for example phenoxyethanol, formaldehyde solution,parabene, pentanediol or sorbic acid as well as those mentioned inenclosure 6, parts A and B of the cosmetic regulation, are furtherclasses of substances. As insect repellents N,N-diethyl-m-toluamide,1,2-pentanediol or insect repellent 3535 come into question, as selftanning agent dihydroxyaceton is suited.

As perfume oils mixtures of natural and synthetic scent substancesshould be mentioned. Natural scent substances are extracts of flowers(lilies, lavendel, roses, jasmin, neroli, ylang-ylang), stems and blades(geranium, patchouli, petitgrain), fruits (anis, coriander, caraway,juniper), fruit shells (bergamot, lemon, orange), roots (macis,angelica, celery, kardamon, costus, iris, calmus), wood (stone pine,sandel, guajac, cedar, rosewood), herbs and grass (tarragon, lemongrass,sage, thyme), needles and twigs (spruce, fir, pine, traipsed), resinsand balsams (galbanum, elemi, benzoe, myrrh, olibanum, opoponax). Rawmaterials from animals are also possible, such as for example zibet andcastoreum. Typical synthetic odour compounds are products from types ofesters, ethers, aldehydes, ketones, alcohols and hydrocarbons. Odourcompounds from types of esters are e.g. benzyl acetate, phenoxyethylisobutyrate, p-tert.-butylcyclohexyl acetate, linalyl acetate,dimethylbenzylcarbinyl acetate, phenylethyl acetate, linalyl benzoate,benzyl formate, ethylmethylphenyl glycinate, allylcyclohexyl propionate,styrallyl propionate and benzyl salicylate. Benzylethyl ether belongsfor example to the ethers, to the aldehydes e.g. the linear alkanaleswith 8 to 18 carbon atoms, citral, citronellal, citronellyloxyacetaldehyde, cyclamen aldehyde, hydroxy citronellal, lilial andbourgeonal, to the ketones e.g. the ionones, α-isomethyl ionon andmethylcedryl ketone, to the alcohols anethol, citronellol, eugenol,isoeugenol, geraniol, linalool, phenylethyl alcohol and terpineol; tothe hydrocarbons mainly the terpenes and balsams belong. However,mixtures of different odour substances are preferred, which togethergive a pleasant smell. Also etheral oils with low volatility, whichoften are used as aroma components, are suited as perfume oils, e.g.sage oil, chamomile oil, carnation oil, melissa oil, mint oil, cinnamonleaf oil, limeflower oil, juniper berry oil, vetiver oil, oliban oil,galbanum oil, labolanum oil and lavandin oil. Preferably used arebergamot oil, dihydromyrcenol, lilial, lyral, citronellol, phenylethylalcohol, α-hexylcinnamon aldehyde, geraniol, benzylaceton, cyclamenaldehyde, linalool, boisambrene forte, ambroxane, indol, hedione,sandelice, lemon oil, mandarin oil, orangenoil, allylamyl glycolate,cyclovertal, lavandine oil, muskateller sage oil, β-damascone, geraniumoil bourbon, cyclohexyl salicylate, vertofix coeur, iso-E-super,fixolide NP, evemyl, iraidein gamma, phenylacetic acid, geranyl acetate,benzyl acetate, rose oxide, romillate, irotyl and floramate, alone or inmixtures.

As colouring agents such substances which are suited and approved forcosmetic purposes can be used, such as for example those mentioned inthe publication “Kosmetische Färbemittel” (cosmetic dyes) of the“Farbstoffkommission der Deutschen Forschungsgemeinschaft”, published byVerlag Chemie, Weinheim, 1984, p. 81-106. These dyes are generally usedin concentrations from 0.001 to 0.1% by weight, based on the wholemixture.

Typical examples of germ inhibiting substances are preservatives withspecific effects against gram-positive bacteria, such as2,4,4′-trichloro-2′-hydroxy diphenylether, chlorohexidin(1,6-di-(4-chlorophenyl-biguanido-hexan) or TCC(3,4,4′-trichlorocarbanilide). Many scent substances and etheral oilsalso have antimicrobial properties. Typical examples are the activeagents eugenol, menthol and thymol in carnation, mint and thyme oil. Aninteresting natural deo substance is the terpene alcohol famesol(3,7,11-trimethyl-2,6,10-dodecatrien-1-ol), which is present in limeflower oil and has a smell of lilies of the valley. Also, glycerolmonolaurate have been used as bacteriostaticum. Normally the content ofthe further germ inhibiting agent is about 0.1 to 2% by weight—based onthe solids content of the preparations.

The cumulative contents of the auxiliary and additional agents can be 1to 50, preferably 5 to 40% by weight, based on the agents. Themanufacture of the agents can take place by common cold or hotprocesses; preferably the work is carried out according to the phaseinversion temperature method.

EXAMPLES

By way of example, the following experiments demonstrate that theβ-glucans of the invention work as active agents for combating theformation of wrinkles in the skin as well as epithelial tissue diseases,e.g. ulcers. The following examples are meant to illustrate theinvention. They are not intended to limit the scope of the invention inany manner or to any degree.

Example I

A panel consisting of 15 female probands aged between 35 and 50 yearswere during a time period of 28 days daily exposed to a daily expositionof different glucans. For this purpose O/W skin cremes with thecomposition stated in table 1 was made by mixing of the phases I and IIat 95° C. TABLE 1 Composition of O/W Skin Cremes Composition Phase IPhase II Cetylstearyl alcohol  8.0 — Ceteareth-12  1.5 — Ceteareth-20 1.5 — Cetearyl isononanoate 15.0 Paraffin oil, viscous  5.0 Baysilonoil M 300  5.0 Glucan 20.0 Glycerol 6.0 Water — 38.0 Sum 36.0 64.0

The probands used the skin cremes daily before going to bed. Withintervals of 7 days the number, depth and length of the skin wrinkleswere determined for each of the participants by means of profilometry ofa selected part of the skin, i.e. a vertcal stripe of 2 cm width and 5cm length, having an upper left and right boundary. which occurs if fromthe nose root a horizontal line is drawn, from this and against theright eye 2, respectively 4 cm, are cleared away and both resultingpoints in each case are elongated in an angle of 270° in each case 2 cm.The dimensionless product of depth, number and length of the skinwrinkles on the day before the beginning of the exposure was set asstandard (=100%), and all the following measurements were related tothis. At the same time the skin roughness of the pro-bands was evaluatedon a scale from 0=“unchanged” to 3 =“strongly improved”. The results aresummarized in table 2. Example 1 is according to the invention, theexamples V1 to V7 are for comparison. It can be seen that the watersoluble β-(1,3) glucans of the invention, which have no (1,6) linkages,exhibit a better effect than known glucans according to the state of theart. TABLE 2 Skin aging and Skin Roughness Skin aging after [d] in [%]Skin roughness after [d] Ex. Glucan 0 7 14 21 28 0 7 14 21 28 1 β-1,3Glucan 100 96 91 85 79 0 1 2 3 3 V1 Schizophyllan¹⁾ 100 98 95 90 86 0 01 2 3 V2 Krestin³⁾ 100 99 96 92 88 0 0 0 1 2 V3 Sclereoglucan⁴⁾ 100 10099 96 88 0 0 0 1 2 V4 CM-Glucan⁵⁾ 100 100 99 96 95 0 0 0 1 1 V5Mannozym⁶⁾ 100 100 100 99 98. 0 0 0 1 1 V6 Lichenin⁷⁾ 100 100 100 100 980 0 0 0 1 V7 Isoiichenin⁸⁾ 100 100 100 100 98 0 0 0 0 1Legend:¹⁾SBG, Biotec Pharmacon ASA, Tromsø²⁾β-(1,3)-/β-(1,6) Glucan, extact from Schizophyllum Commune

Example II

A limited pilot clinical study examining the ability of the abovedescribed soluble beta-1,3/1,6-glucan to facilitate healing of diabeticulcers was initiated. Fifteen patients with type 1 (1 patient) or type 2(14 patients) diabetes were included (for inclusion and exclusioncriteria, see Table 3). The single patient with type 2 diabetes wasplaced in the control group. All patients had clinically typical“neuropathic” ulcers of the lower extremities (stage II and III WHOcriteria). SBG refers to the soluble beta glucan of this invention.TABLE 3 Pilot study with topical SBG treatment of chronic diabeticulcers Inclusion criteria: Type 1 or type 2 diabetes mellitus,complicated by chronic ulcers of the lower extremities (neuropathictype, 1-40 cm² after initial debridement; infection and cellulitescontrolled before randomisation) Adequate arterial circulation (normalmalleolus-brachial indices as determined by Doppler techniques) Signedinformed consent Exclusion criteria: Osteomyelitis of affected areaUlcer after debridement <1 cm² or >40 cm² or sum of all ulcers >100 cm²Patients with ulcers of other than diabetic etiology GangreneConcomitant disease requiring radiotherapy, chemotherapy,immunosuppresive medications, corticosteroids, connective tissuedisorder, cancer Pregnant or lactating women

Six patients received conventional therapy: diabetes control (systemicinsulin, peroral sulfonyl urea), off-loading of pressure from theaffected area; debridement and local application of antibacteriallevamicol (0.75%). These six patients constituted the control group.Nine patients received similar diabetic control and debridement as thecontrol group, but topical application of a 2% solution of solublebeta-1,3/1,6-glucan instead of levamicol (for details of treatment, seeTable 4). TABLE 4 Pilot study with topical SBG treatment of chronicdiabetic ulcers Treatment Treatment was given every second day in theout-patient setting and every day for hospitalized patients as follows:Surgical debridement, removal of necrotic and hyperkeratotic tissuesUlcers were washed with 0.9% NaCl Soluble beta-1,3/1,6-glucan as 2%solution (in aq.pur.) or Levamicol (0.75% levamicatin inpolyethyleneoxide) treatment: Dose = ulcer volume. In case of deepulcers, a “gauzeball” was inserted. Covering with gauze bandages, etc

All patients that received beta-glucan treatment had complete healing ofulcers within 100 days and most ulcers healed by day 35 (see FIG. 1).

In contrast, only 2 (of 6 eligible) patients in the control groupachieved complete closure of ulcers and these lesions were both small(<2 cm2). In one control case, beta-glucan treatment had to beintroduced after 2 weeks for ethical reasons (see asterisk, FIG. 1).

Example III

Herein is described the healing of aphthous ulcers in a 62 year oldmale, with a history of suffering from recurrent aphthous ulcers on themucosal epithelial layer in the oral cavity during several years withreappearance every 3-12 months lasting for several weeks. The ulcermanifested as 2-6 mm first red edged and subsequently as white edgedcraters particularly under the tongue, but also elsewhere in the oralcavity. The patient started using 10 mL of a 10 mg/mL SBG solution as anoral rinse every second day during outbreak of the ulcers, resulting incomplete healing of the ulcers during the first week of treatment. Thepatient has for three years used SBG as weekly rinse, and has since thestart of treatment not suffered from recurrence of aphthous ulcers.

1. A method of treating an ulcer comprising applying to the ulcer apreparation comprising water-soluble β-(1,3) glucan with β-(1,6) linkedside-chains, where said side-chains comprise β-(1,3) linkages or up tofour consecutive β-(1,6) linkages, as active substances.
 2. The methodaccording to claim 1, wherein the glucans are used in amounts of 0.1% to25% by weight based on the preparation.
 3. The method of claim 1,wherein the ulcer is chronic ulcer.
 4. The method of claim 3, whereinthe chronic ulcer is diabetic ulcer.
 5. The method of claim 3, whereinthe chronic ulcer is aphthous ulcer.
 6. A glucan for use in atherapeutic composition for treatment of ulcers, wherein said glucancomprises water-soluble β-(1,3) glucan with β-(1,6) linked side-chains,where said side-chains comprise β-(1,3) linkages or up to 4 consecutiveβ-(1,6) linkages.
 7. The glucan of claim 6, wherein the said glucans arebased on yeast of the family Saccharomyces.
 8. The glucan of claim 6,wherein the said glucans are obtained by contacting glucans with β-(1,3)and β-(1,6) linkages with β-(1,6) glucanases.
 9. The glucan of claim 8,wherein said glucanases are based on Trichoderma harzianum.
 10. Theglucan of claim 6, wherein the said glucans are used in amounts of 0.1%to 25% by weight relative to the therapeutic composition.
 11. The glucanof claim 6, wherein the ulcer is chronic ulcer.
 12. The glucan of claim11, wherein the chronic ulcer is diabetes ulcer.
 13. The glucan of claim11, wherein the chronic ulcer is aphthous ulcer.